The isolation of complex macromolecular assemblies at the concentrations required for structural analysis represents a major experimental challenge. Here we present a method that combines the genetic power of site-specific recombination in order to selectively "tag" one or more components of a protein complex with affinity-based rapid filtration and a final step of capillary-based enrichment. This modified form of tandem affinity purification produces highly purified protein complexes at high concentrations in a highly efficient manner. The application of the method is demonstrated for the yeast Arp2/3 heptameric protein complex involved in mediating reorganization of the actin cytoskeleton.
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http://dx.doi.org/10.1016/j.ab.2013.03.035 | DOI Listing |
Front Pharmacol
January 2025
Department of Pharmacognosy, School of Pharmacy, Xinjiang Medical University, Urumqi, China.
Objective: To elucidate the metabolic mechanisms by which acteoside (ACT) isolated from alleviates cancer-related fatigue (CRF) in a murine model of colon cancer with cachexia.
Methods: BALB/c mice inoculated with C26 colon cancer cells were treated with paclitaxel (PTX, 10 mg/kg) and ACT (100 mg/kg) alone or in combination for 21 days. Fatigue-associated behaviors, tumor inhibition rate, and skeletal muscle morphology assessed by hematoxylin-eosin (H&E) staining and electron microscopy were evaluated.
J Chromatogr Sci
January 2025
Department of Chemistry & Biochemistry, Ohio University, Athens, OH, USA.
The valid method was developed for analyzing empagliflozin in serum/plasma/urine using a molecularly imprinted ghost polymer-solid-phase extraction approach (MISPE) with liquid chromatographic methodology. Methacrylic acid (MAA) was used as the monomer, 2,2 azobis isobutyronitrile as the initiator and ethylene glycol dimethacrylate as the cross-linker in the free radical polymerization procedure. Empagliflozin was loaded onto the polymer and eluted with 1 mL of a 9:1 MeOH:acetic acid solution.
View Article and Find Full Text PDFBMC Complement Med Ther
January 2025
Henan Institute of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou, China.
Background: Amorpha fruticosa was used for treating burn, ambustion, carbuncle, and eczema in the traditional Chinese medicine. Although more and more attention has been paid to its biological activity recently, the antitumor activities of the essential oils (EOs) extracted from its leaves (AFLEO) and flowers (AFFEO), and their molecular mechanisms have never been reported up to now. The objective of present study was to examine the chemical compositions of AFLEO and AFFEO, then investigate the effects and pharmacological mechanism of EOs against hepatocellular carcinoma (HCC).
View Article and Find Full Text PDFSci Rep
January 2025
Johnson & Johnson, Therapeutics Discovery, Spring House, PA, USA.
Solution-based affinity assays are used for the selection and characterization of proteins that could be developed into therapeutic molecules. However, these assays have limitations for cell-surface proteins as in most cases their purification requires detergent solubilization and are unlikely to assume conformations in solution that resemble their native states in cell membranes. This report describes a novel electrochemiluminescence-based method, called MSD-CAT, for the affinity analysis of antibodies binding to cell-surface receptors.
View Article and Find Full Text PDFJ Hazard Mater
January 2025
Key Laboratory for Green Chemical Process of Ministry of Education, Hubei Key Lab of Novel Reaction & Green Chemical Technology, School of Chemical Engineering and Pharmacy, Wuhan Institute of Technology, Wuhan 430205, PR China. Electronic address:
Pd-zeolite is considered one of the most promising passive NO adsorber (PNA) materials for NO purification in diesel vehicles during cold start. Nevertheless, the scarcity and high cost of the precious metal Pd restrict the industrialisation of Pd-zeolites as PNA. This work developed a bimetallic Mn and Ba co-modified SSZ-13 as non-precious metal PNA material.
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